TUBright: A Peptide Probe for Imaging Microtubules,Analytical Chemistry

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TUBright: A Peptide Probe for Imaging Microtubules
Analytical Chemistry ( IF 6.7 ) Pub Date : 2022-08-02 , DOI: 10.1021/acs.analchem.2c01285
Songbo Xie ₁ , Jingrui Li ₁ , Shuang Sun ₁ , Wei Chen ₂ , Haisu Cheng ₂ , Yinlong Song ₂ , Yuyang Li ₁ , Min Liu ₁ , Xueliang Zhu ₃ , Xin Liang ₂ , Jun Zhou _{1,

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Affiliation

In vitro assays using reconstituted microtubules have provided molecular insights into the principles of microtubule dynamics and the roles of microtubule-associated proteins. Emerging questions that further uncover the complexity in microtubule dynamics, especially those on tubulin isotypes and post-translational modifications, raise new technical challenges on how to visualize microtubules composed of tubulin purified from limited sources, primarily due to the low efficiency of the conventional tubulin labeling protocol. Here, we develop a peptide probe, termed TUBright, that labels in vitro reconstituted microtubules. TUBright, when coupled with different fluorescent dyes, provides flexible labeling of microtubules with a high signal-to-noise ratio. TUBright does not interfere with the dynamic behaviors of microtubules and microtubule-associated proteins. Therefore, TUBright is a useful tool for imaging microtubules, making it feasible to use tubulin from limited sources for answering many open questions on microtubule dynamics.

中文翻译：

TUBright：一种用于微管成像的肽探针

使用重组微管的体外测定为微管动力学原理和微管相关蛋白的作用提供了分子见解。新出现的问题进一步揭示了微管动力学的复杂性，特别是关于微管蛋白同种型和翻译后修饰的问题，对如何可视化由有限来源纯化的微管蛋白组成的微管提出了新的技术挑战，这主要是由于传统微管蛋白标记的低效率协议。在这里，我们开发了一种肽探针，称为 TUBright，可在体外进行标记重组微管。TUBright 与不同的荧光染料结合使用时，可以灵活地标记具有高信噪比的微管。TUBright 不会干扰微管和微管相关蛋白的动态行为。因此，TUBright 是一种有用的微管成像工具，使得使用有限来源的微管蛋白来回答许多关于微管动力学的开放性问题变得可行。

更新日期：2022-08-02

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